The present invention relates to compositions and methods of use for the treatment of bacterial, fungal, and viral infections of the dermis and cuticle. More specifically, the present invention relates to compositions and methods of use of Emu Oil, and it various associated fractions, in combination with the appropriate medicaments, in the treatment of bacterial, fungal, and viral infections of the dermis and cuticle. The present invention also relates to the utilization of therapeutic compositions comprised of Emu Oil in combination with a probiotic, viable Bacillus bacteria, spores, and extracellular supernatant products, as well as the extracellular product of Pseudomonas lindbergii, as a topical agent for the prevention and/or control of infections caused by bacterium, fungi, yeast, and virus, and combinations thereof.
1. Emu Oil
Emu Oil, is an animal-derived lipid composition, extracted from the Emu (Dromais Novae-Hollandiae), a flightless bird part of a group called ratites (which also includes the ostrich and the kiwi), indigenous to Australia and New Zealand.
Emu Oil is extracted from a thick fat-pad on the back of the bird which putatively functions to protect the animal from the extreme temperatures in its Australian homeland. The fat is carefully extracted to prevent the formation of trans-fatty acids, wherein approximately 100 pounds of fat produces approximately 50 to 90 pounds of unrefined, pale yellow oil. The chemical composition and characteristics of Emu Oil has been quantitatively ascertained and is set forth below in Table I.
As illustrated in Table I, when correctly extracted and processed, Emu Oil is comprised of approximately 50% to 70% monounsaturated fatty acids, with the rest being both saturated and polyunsaturated fatty acids (see e.g., American Emu Association News, March 1995). Emu Oil is almost purely triglyceride in nature, which makes it an almost completely neutral lipid. In addition, the monounsaturated fatty acid, oleic acid, is the largest single fatty acid component of Emu Oil. Traditional beliefs of geographically widely-separated Australian Aboriginal communities agree on the beneficial properties of Emu Oil as a natural remedy. The oral history of the Australian Aborigines indicates their use of Emu Oil for over forty thousand years to reduce pain and stiffness in sore muscles and joints, to help expedite wound healing, as a dermal protectorate from the effects of wind and sun, and in the treatment of bruised subcutaneous tissue, burns and dry skin problems. Methods of administration are quire varied. For example, Aborigines have revealed methods of treatment which included hanging an Emu skin on a tree to collect the oil, and wrapping the affected area on the individual in a freshly-killed Emu skin. However, it is believed that in both of the aforementioned scenarios, the catalyst of the suns"" heat was used to liquefy the Emu fat and enhance its absorption qualities.
Documented records of the utilization of Emu Oil may be antedated well over 100 years (see e.g., Whitehouse, et al., 1996. Concerning Emu Oil and its anti-arthritic activity. Fifth Queensland Poultry Science Symposium, Gatton College). The use of Emu Oil was among many natural remedies adopted by settlers from the original inhabitants of Australia. The first report known was published in the Australian Post regarding experiments by Dr. Peter Gosh (Raymond Purves Bone and Joint Research Laboratories, University of Sydney at the Royal North Shore Hospital) and Dr. Michael Whitehouse (Department of Pathology, University of Adelaide), wherein the Emu Oil was required to be massaged vigorously onto the sore muscle or joint and the process repeated as often as required, hence pressure, heat and duration of rubbing were all deemed to be relevant factors.
This, although Emu Oil has been previously described, the majority of its uses or properties/characteristics is anecdotal in nature. These uses and properties include (see e.g., DuBois, 1999. Explore Issue 8:1-10): (i) its ability to act as a dermal penetrant and medicament carrier; (ii) its anti-inflammatory properties; (iii) its ability to act as an emollient/emulsifier; (iv) its bacteriostatic properties; (v) its low potential for irritation of the skin; (vi) its non-comedogenic properties (i.e., it does not clog up pores); and (vii) its moisturizing, wound-healing, general xe2x80x9canti-agingxe2x80x9d properties. However, the quantitative information currently available almost exclusively relates to the benefits of Emu Oil as an anti-inflammatory agent for arthritis, its uses for cardiovascular health when ingested, which is similar to the use of Omega-3 fish oils to improve high-density lipoprotein (HDL) cholesterol, and its moisturizing and general xe2x80x9canti-agingxe2x80x9d properties.
There is much anecdotal material available on the anti-inflammatory abilities of Emu Oil. It has been shown to reduce pain, swelling and stiffness in joints, to reduce recent bruising and muscle pain, and ease sports related muscle strains as well. Studies have shown that different Emu Oils (i.e., oils which were extracted by different methodologies, from different sources, and the like) possessed different levels of anti-inflammatory ability. The ability of Emu Oil to penetrate the stratum coneum dermal barrier and concomitantly act as a carrier, makes it highly valuable for use in therapeutic compounds in the prevention and/or treatment of a variety of conditions. This ability is believed to be primarily due to both its extremely high content of oleic acid and a total lack of indigenous phospholipids. Accordingly, Emu Oil could be combined with various medicinals or cosmetic materials to facilitate their ability to penetrate this layer of keritinized tissue in a more efficacious and cost-effective manner than the currently-utilized liposome- and iontophorisis-based technologies. For example, the ability of Emu Oil to act as a trans-dermal penetrant with respect to Ketoprofen, a well known non-steroidal, anti-inflammatory drug (NSAID) found in Actron(trademark) and like products, was examined in a recent study performed at Auburn University. Ketoprofen is one of the proprionic acid derivative drugs, which have been utilized in numerous European countries for more than 15 years as an effective treatment for rheumatoid arthritis and osteoarthritis. Although it is available in more than 80 countries throughout the world, it did not receive approval for over-the-counter (OTC) use in the United States until 1996. Although Ketoprofen is readily absorbed, it frequently produces a number of adverse side-effects in the gastrointestinal tract when taken orally. Moreover, the oral administration of Ketoprofen has also been associated with such serious deleterious physiological side-effects as renal dysfunction, marked edema, and hepatic dysfunction (e.g., jaundice). The utilization of a topically-administered Ketoprofen composition to the dermis over the inflamed tissues or joints would perhaps mitigate some of the aforementioned side-effects and may also potentially result in the accumulation of the drug within associated synovial tissues, the site of the desired anti-inflammatory reaction. However, recent studies in which Ketoprofen was topically-administered without the utilization of dermal-penetrants (e.g., Emu Oil) demonstrated that this compound was adsorbed through viable, keritinized dermal tissue in a very limited concentration, if at all.
Conversely, the results demonstrated that the concomitant utilization of a dermal-penetrant produced markedly elevated adsorption of the compound. Specifically, an Emu Oilxe2x80x94propanolxe2x80x94Ketoprofen combination was shown to produce a 3-times higher serum levels in mice following trans-dermal application, than either a DMSOxe2x80x94Bovine Serumxe2x80x94Ketoprofen or an Isopropyl alcoholxe2x80x94Ketoprofen combination. This result was particularly encouraging due to the fact that Emu Oil was approved by the FDA for human use in July of 1992, and DMSO has not yet received such approval.
In a related study, the ability of Emu Oil to decrease the concentration of inflammatory molecules was examined (see Smith and Craig-Schmidt, AEA Convention Las Vegas, Nev. (Jun. 6-8, 1995)). Eicosanoids are hormone-like compounds synthesized from essential fatty acids and have been demonstrated to be synthesized in dermal tissue (see e.g., Wilkerson and Walsh, 1977. J. Invest. Dermatol. 68: 210-214). While some of these compounds serve normal physiological functions, others are involved in the inflammatory response. In this study, prostaglandin F2a (PGF2a.) was utilized as an indicator of ecosanoid synthesis within the dermal tissue. The topical administration of was shown to decrease ecosanoid production in skin, as reflected by suppression of PGF2a. This result may offer a possible biochemical explanation for the reported beneficial effects of topically administered Emu Oil.
Additionally, in 1995, Australian researchers isolated a component in Emu Oil which appears to be at least one of the active ingredients directly responsible the oil""s anti inflammatory activity. Thus, this substance could potentially be utilized to develop or isolate additional anti inflammatory medications which are without deleterious physiological side-effects, are non-irritating, which possess long-term biological and physiological activity, and which are far less expensive than currently-utilized anti-inflammatory regimens.
Emu Oil also possesses a high degree of emollient/emulsification properties, and hence has good xe2x80x9cblendabilityxe2x80x9d. In practice this means that Emu Oil has the ability to blend or make oil and water misable, producing a cream that does not feel oily on the skin. One inherent problem is that most creams do not penetrate the dermal barrier, however this is ameliorated by the utilization of Emu Oil without leaving an oily residue behind. This bodes very well for its future use in both the cosmetic and pharmaceutical industries.
An additional property of Emu Oil is that it is bacteriostatic. Recent studies have demonstrated that in its pure state, Emu Oil grows no bacterial organisms. Thus, pure non-contaminated Emu Oil has a long shelf-life due to its bacteriostatic nature and due to its low levels of polyunsaturated fats which are the most subject to oxidation and eventual rancidity. Similarly, Emu Oil""s bacteriostatic activity will be of useful in both cosmetic and pharmaceutical industries.
Emu Oil also possesses an extremely low potential for irritation of the skin. Moreover, it has also been shown to have almost no side-effects, which means that (even at full strength), Emu Oil has irritation levels so low that they are the same as those found in putting water on the skin (i.e., is practically nonexistent). This characteristic is unusual, as most anti-inflammatory drugs are irritating, when applied topically, and possess side-effects.
Emu Oil is non-comedogenic in nature, and does not xe2x80x9cclogxe2x80x9d the pores of the skin nor tend to cause acne when used. This tendency cannot be said for, e.g., mineral oil (which is one of the current, popular carrier oils in cosmetics and rubbing oils) which frequently causes outbreaks of acne when used.
Finally, Emu Oil is a highly efficacious moisturizing agent, which adds to its protective ability and promotes anti-aging of the skin. Researchers believe that its unique combination of saturated and unsaturated fatty acids may be an explanation for its ability to enhance the willingness of the upper layers of the skin to retain water. For example, application of Emu Oil has been demonstrated to increase the overall thickness of human skin by approximately 2.5-times, thus reducing its tendency to form xe2x80x9cwrinklesxe2x80x9d. In addition, there is much anecdotal information regarding the anti-aging and wound healing abilities of Emu Oil. A double-blind study is currently being performed at the Timothy J. Harner Burn Center (affiliated with the University Medical Center in Lubbock, Tex.) to authenticate this anecdotal material.
The general xe2x80x9canti-agingxe2x80x9d properties of Emu Oil was examined at the Boston University School of Medicine. In this double-blind study, a refined Emu Oil known as Kalaya (New World Technology; Los Angeles, Calif.) was topically-administered daily to depilated mice, over a two-week time-period. Corn oil was utilized as the negative control substance. Results demonstrated that the refined Emu Oil produced a 20% increase in the overall rate of DNA synthesis within the skin cells of these animals, whereas the rate of DNA synthesis within the negative control animals remained normal. A marked increase in the overall thickness of the skin, to which the Emu Oil had been applied, was also found. In addition, over 80% of hair follicles which were quiescent at the time of the initiation of the study, were stimulated by the application of the Emu Oil and began to produce a viable hair shaft. Typically, hair follicles go through stages from a quiescent phase, to an active hair-growth phase, and back to the quiescent phase again. The administration of Emu Oil was found to not only stimulate the hair follicles into the active phase, but it kept them in this phase during the entire period of administration, as well.
Studies regarding the properties of Emu Oil have expanded to prominent noted facilities/groups including, but not limited to: Auburn University; The Arthritis Clinic, Ardmore, Okla.; Texas Technical University; Timothy J. Harnar Burn Center; and Iowa State University.
The use of Emu Oil in veterinary medicine has also gained favor (see e.g., Zimmer, 1999. J. Equine Med. 56: 112-117). Emu Oil is frequently used in combination with DMSO or dexamethasone, or Gentamicin for the management of wounds. The treatment of non-suturable wounds (e.g., distal leg wounds where there is decreased muscle mass), by twice-daily application of Emu Oil was shown to markedly increase epithiliazation of these wounds, while concomitantly reducing the development of necrotic tissue and scarring. Similarly, the frequency of dehiscence of sutured wounds was also demonstrated to be markedly reduced in Emu Oil-treated equines. Emu Oil in combination with NSAID is also used to control stiffness and pain in those affected joints in lame or arthritic horses. A frequent winter lesion seen in dairy cattle is frosted teat ends, wherein the teat end freezes and skin around the teat sloughs. Topical administration of Emu Oil has been found to accelerate the healing process and allows the continued milking of the cow during this process. The bacteriostatic properties of Emu Oil is also effective in the prevention and/or treatment of infections of the teat in dairy cows due to milk residues. Similarly, Emu Oil is more effective in the treatment of ringworm lesions (commonly seen in calves) than other conventional techniques (e.g., bleach, iodine preparations, and the like). Another area in which Emu Oil is utilized in veterinary medicine is the treatment of lesions or sores caused by casts. When a cast area is applied it frequently retains moisture or causes pressure on bony protuberances, resulting in the formation of dermatitis or cast sores. Following the removal of the cast, the use of Emu Oil greatly accelerates the healing process of these aforementioned sores.
2. Dermal Infections
Dermal infections, especially those caused by mycotic pathogens, make-up a considerable percentage of the sale of prescription and over-the-counter medications that are sold annually worldwide. According to the Center for Disease Control and Prevention (CDCP), there is currently a dramatic rise in the number of reported mycotic and bacterial skin infections. Annual sales of dermal and cuticular anti-fungal agents is currently exceeding two billion U.S. dollars each year. Moreover, dermal mycotic illness was recently shown to be increasing at a rate of approximately 9% to 15% per annum, depending upon the specific pathogen and disease. One of the primary factors responsible for the growth of these markets is the fact that more fungal pathogens are becoming resistant to the commonly-utilized anti-fungal agents each year. Examples of anti-fungal agents which are commonly-utilized, include, but are not limited to: Fluconazole (Diflucan(copyright); Pfizer Pharmaceutical), Intraconazole (Sporonox(copyright); Janssen Pharmaceutical), Miconazole Nitrate, Ketoconazole, Tolnaftate, Lamasil, Griseofulvin, Amphotercin B, and other compounds and the formulations thereof.
New generations of anti-fungal and anti-bacterial drugs and preparations are being developed every year to replace those medication in which pathogens have become resistant. As the search for more effective anti-microbial agents continues, so does the search for xe2x80x9ccarrying agentsxe2x80x9d which are utilized to disperse and facilitate penetration of these medications through the various dermal and cuticular membranes and tissues. However, to date there has been little success in finding an agent that is able to penetrate dense cuticular material such as finger/toenails and animal hooves.
Diseases that are most common to human dermal and cuticular membranes include: (i) Candidaiasis (e.g., caused by Candida albicans, Candida tropicalis, Candida golbratta, Candida parapsilosis); (ii) Tineal diseases, also known as Athletes Foot (Tinea Pedis), Jock Itch (Tinea Cruis), Scalp Infection (Tinea Capitis), Ring Worm, and Beard infections (Tinea Barbae), are all caused by the Trichophyton species, including, but not limited to: Trichophyton mentagrophytes; (iii) diseases which are caused by bacterial pathogens, including, but not limited to: Pseudomonas aeruginosa, Staphylococcus aerues, Staphylococcus epidermidus, and Propionibacterium acnes; and (iv) diseases which are caused by viral pathogens, including, but not limited to: Herpes simplex I and II, and Herpes zoster. Perhaps one of the most difficult-to-treat diseases of fungal etiology are fungal infections of the toenail or fingernail (i.e., Onychomycosis) due to the inability of the currently-available therapeutic compositions to penetrate the dermis or cuticle. The pathogen most commonly associated with this very difficult to treat disease is Trichophyton rubrum. 
In animals, the most common dermal fungal disease is Ring Worm. In animal hooves, especially athletic equine, there are several diseases of the hoof that are potentially quite serious and difficult to treat, including: White Line Disease (also known as xe2x80x9cSeedy Toexe2x80x9d), Hoof Thrush (another yeast- or Candida-related malady), and Drop Sole. In addition, Clubbed Foot is another dermal fungal disease that is of significant concern to the equine industry.
3. Bacillus coagulans 
Bacillus coagulans is a non-pathogenic gram positive spore-forming bacteria that produces L(+) lactic acid (dextrorotatory) in homofermentation. This microorganism has been isolated from natural sources, such as heat-treated soil samples inoculated into nutrient medium (see e.g., Bergey""s Manual of Systemic Bacteriology, Vol. 2, Sneath, P.H.A., et al., eds., (Williams and Wilkins, Baltimore, Md., 1986)). Purified Bacillus coagulans strains have served as a source of various enzymes including, but not limited to: restriction endonucleases (see U.S. Pat. No. 5,200,336); amylase (see U.S. Pat. No. 4,980,180); lactase (see U.S. Pat. No. 4,323,651); and cyclo-malto-dextrin glucano-transferase (see U.S. Pat. No. 5,102,800). Bacillus coagulans has been used to produce lactic acid (see U.S. Pat. No. 5,079,164). In addition, a strain of Bacillus coagulans (designated Lactobacillus sporogenes, Sakaguti and Nakayama (ATCC 31284)) has been combined with other lactic acid-producing bacteria and Bacillus natto to produce a fermented food product from steamed soybeans (see U.S. Pat. No. 4,110,477). Bacillus coagulans strains have also been used as animal feed additives for poultry and livestock to reduce disease and improve feed utilization and to, therefore, increase growth rate in the animals (see PCT Patent Application Nos. WO 9314187 and WO 9411492).